Hematology

What is Karl Landsteiner credited

with discovering?

Blood
Compositio
n

Key Terms
Anticoagulant: an agent that prevents
the clotting of blood.
Examples are EDTA, Citrate and Heparin

Capillary: small blood vessel that

connects arterioles and venules
Hematoma: a subcutaneous mass of
blood at a venipuncture site

Key Terms
Hemoglobin: the oxygen carrying
molecule of red blood cells
Hemolysis: the breakdown of red blood
cells, with the release of hemoglobin
into the plasma or serum. Cannot use
hemolyzed samples in lab tests
Icteric: jaundiced; dark yellow or
greenish serum or plasma

Key Terms
Lipemic: having abnormally high level
of fat. Milky looking samples
Plasma: pale yellow part of whole
blood; contains all clotting factors
Serum: liquid portion of blood without
the protein fibrinogen, which is one of
the clotting factors; clot removed

Key Terms
Agglutination: the clumping together
of rbc by the action of an antibody
When A cells are added to a blood sample,
agglutination will occur only in the presence of
anti-A

Serology: the study of antigen

antibody reactions using laboratory
tests

Function of Blood
Transporting fluids such as:

Nutrients from digestive tract

O2 from lungs
Waste from cells
Hormones

Aids in heat distribution

Regulates acid-base balance

Composition of Blood
Plasma: liquid portion of blood w/out
cells
Contains all of the following
Water
Electrolytes

Nutrients
Metabolic waste
product
Hormones
Vitamins and enzymes
Plasma proteins such as fibrinogen, albumin and
globulin

Composition of Blood:
Erythrocytes
Red blood cells are responsible for:
Transport of oxygen and nutrients
Removal of waste and CO2 from the cells
Distribution of heat

Hemoglobin: the O2 carrying potential

Composition of Blood:
Leukocytes
WBC are responsible for:
Phagocytosis to engulf and absorb waste
material and harmful microorganisms in the
blood stream and tissues
Synthesis of antibody molecules
Inflammation process
Production of heparin component found in
lung and liver tissue which have the ability to
prevent clotting of blood.
Heparin used in the treatment of thrombosis

Cell Morphology

Lymphocyte

rbc

Platelet

Segmented Neutrophil

Composition of Blood:
Leukocytes
Types of Leukocytes
Granulocytes
Neutrophils
Eosinophils
Basophils

Agranulocytes
Lymphocytes
Monocytes

Composition of Blood:
Thrombocytes
Platelets the smallest of the solid
components of the blood
Responsible for the clotting process
Coagulation: term for clotting
Embolism: a blood clot which is
moving through the body

Forensic Characterization
of Blood
1st questions a criminalist has to answer is:
Is it blood
If yes, is it human
If yes, can it be associated w/ a particular person

Preliminary color test for blood is the

Benzidine color test but this is
carcinogenic so phenolphthalein is used
and is known as the Kastle-Meyer Color
test

Forensic Characterization
of Blood
Hemoglobin possesses peroxidase
like activity which when mixed with
phenolphthalein and hydrogen
peroxide it will cause the formation of
a deep pink color
Kastle-Meyer is not specific for blood
as some vegetables such as potatoes
and horseradish contain peroxidase
and can react; however they should
not be common at a crime scene so it
is often considered a good indicator

Forensic Characterization
of Blood
Hemastix strips can also be used to
detect the presence of blood
Luminol test is another presumptive
test for blood which produces light
rather than a color reaction. Objects
being tested must be in a dark
location to view the luminescence
(emission of light).
Extremely sensitive and can detect blood
diluted up to 300,000 times
Luminol will not interfere with other DNA testing

Forensic Characterization
of Blood
Once blood is found it must be
determined to be human
Precipitin test is the standard test
Reagents are available to determine if blood is
dog, cat or deer
A positive test is a cloudy ring or band at the
point where the two liquids meet
Only a small amount of sample is required
Precipitin test is very sensitive and can test
positive on a sample as old as 10 years
Extracted tissue samples from mummies as
old as 4,000 years have tested positive

Forensic Characterization
of Blood
Gel diffusion is another method which
can be used.
Antigen and antibody assay

Gel electrophoresis can also be used

to determine if a sample is human
blood or not
Antigen and antibody assay
Uses electrical current and ppt. line forms where the
two samples meet if it is a positive reaction

Blood Types
Four Major Groups
A

AB

Blood types are inherited from your

parents
Antigen is present on the red blood
cell; typing is done w/rbc
Antibody is present in the plasma;
antibody screening done on plasma

Blood Types
O negative
Universal donor
It carries no antigen

AB positive
Universal recipient
It carries no antibodies in the plasma

43% of population are O, 42% A, 12%

B and 3% AB

Rh Factor (D antigen):
found on the surface of
rbc

Rhesus factor:
discovered in rhesus
monkeys in 1937
Can be
phenotypically
positive or negative
Positive is dominant
over negative
If positive is present,
then you will express
positive phenotype

+
_

++

+-

+-

--

+- +-

+- +-

Blood Types: ABO

Controlled by
GENETICS!!!!!

AB

BO

Two Categories

AO

OO

ABO and Rh
A
A
A
O

AA AO
AO OO

AA AB

AB BB

Blood Types: What Ag do

they have?
A positive blood has which Ag present
on rbc?
A antigen and Rh antigen

B negative blood has which Ag

present on rbc?
B antigen

AB + blood type has which Ag

present?
Both A and B Ag and Rh antigen on the rbc

Who can donate to whom?

A + donor: what blood types can this
pt. receive?
A+, A-, O+, O-

Your blood type is B-, what blood

types can donate to you?
B- and O-

AB- as a last resort

ABO Blood Types

Agglutination Reactions

Anti-A

Anti- B

Anti-A &
Anti B

Yes

No

Yes

No

Yes

Yes

AB

Yes

Yes

Yes

No

No

No

Examples of ABO blood typing

O negative carries no Ag and therefore does not react with any Anti A,
B, AB

Pos reaction

Neg reaction

Rh Antigen and Antibody Interaction

Anti- Rh
Rh +

YES

Rh -

NO

Normal Ranges
RBC:

female 3.6-5.0x106mm3
male 4.2-5.4x106mm3

WBC:

4.5-10.5x103mm3 (African Americans is sltly

lower 3.2 is still normal)

HCT:

female 36-48%

Hgb:

female 12-16 g/dL male 14-17.4 g/dL

male 42-52%

Platelets: 140-400x103mm3

Blood Cell
Maturation

Blood Splatter Analysis

Location, distribution, and
appearance of blood stains are an
important part of forensics
Investigators try to determine:
Direction
Dropping distance
Angle of impact

Splatter analysis is often used for

crime scene reconstruction

Blood Splatter Analysis

Factors which influence stain patterns
are:
Surface texture
Direction of travel
Pointed end of bloodstain always faces its direction of
travel

Angle of impact is determined by measuring the

degree of circular distortion of the stain
Blood striking a surface at right angles gives rise to a
nearly circular stain
As the angle decreases, the stain becomes elongated
in shape

DNA

DeoxyriboNucleic Acid
Found in the nucleus
46 chromosomes
25,000 genes
Structure determined by Crick and
Watson
DNA fingerprinting by Alec Jeffreys

Structure of DNA
A polymer made of repeating
nucleotides
Nucleotide consists of a deoxyribose
sugar, a phosphate, and a nitrogen
base (Adenine, Thymine, Cytosine, or
Guanine)
Double stranded, helical
Complementary base
pairing, A=T, G=C

DNA at Work
DNA in nucleus is copied into a strand
of RNA (transcription)
RNA is read at the ribosome to make
assemble amino acids into proteins
(translation)
Every 3 bases on DNA
codes for a different
amino acid

Replication of DNA
Replication the synthesis of new
DNA from existing DNA in the nucleus
DNA polymerase assembles
new DNA strand and proofreads it
Replication occurs in nucleus prior to
cell division

Polymerase Chain
Reaction
A technique for replication, or
amplifying, a portion of DNA outside
the cell
Each cycle doubles
the number of copies
1
1x107 in 30 cycles

DNA Typing with Tandem

Repeats
Region of chromosome that contains multiple
copies of a core DNA sequence arranging in a
repeating fashion between the coding regions
(genes)
Restriction Fragment Length Polymorphisms used
enzymes to cut the DNA around these tandem
repeat sites and then run them on a gel
electrophoresis
A Southern blot was then performed and
radioactive probes were hybridized to help
visualize the RFLPs

RFLPs

PCR
PCR has the following advantages:
1. PCR can use shorter sequences
2. shorter pieces more stable
3. smaller amounts of DNA can
be used (10-9 gram)

Short Tandem Repeats

(STRs)
A region of a DNA molecule that
contains short segments of 3-7
repeating base pairs.
Generally less than 450 bp long
Less degradation
Can be PCRd
Can multiplex a large number of
these STRs at once
US uses 13 STRs for tests

 Capillary electrophoresis
Sex Identification by focusing on the
amelogenin gene